1a. How is biological sample prepared for the insertion into the transmission electron microscope for single...

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Biology

1a. How is biological sample prepared for the insertion into thetransmission electron microscope for single particle averaging? b.The single image of a molecule in cryo-EM is very noisy. What isdone to improve the clarity of the image? What does that mean interms of statistics?

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1a Sample preparation in TEM transmission electron microscope can be a complex procedure TEM specimens should be less than 100 nanometers thick for a conventional TEM Unlike neutron or XRay radiation the electrons in the beam interact readily with the sample an effect that increases roughly with atomic number squared Z2 High quality samples will have a thickness that is comparable to the mean free path of the electrons that travel through the samples which may be only a few tens of nanometers Preparation of TEM specimens is specific to the material under analysis and the type of information to be obtained from the specimen Materials that have dimensions small enough to be electron transparent such as powdered substances small organisms viruses or nanotubes can be quickly prepared by the deposition of a dilute sample containing the specimen onto films on support grids Biological specimens may be embedded in resin to withstand the high vacuum in the sample chamber and to enable cutting tissue into electron transparent thin sections The biological sample can be stained using either a negative staining material such as uranyl acetatefor bacteria and viruses or in the case of embedded sections the specimen may be stained with heavy metals including osmium tetroxide Alternately samples may be held at liquid nitrogen temperatures after embedding in vitreous ice In material science See Answer

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